Kumai Yasuhiro, Ooboshi Hiroaki, Kitazono Takanari, Takada Junichi, Ibayashi Setsuro, Fujishima Masatoshi, Iida Mitsuo
Department of Medicine and Clinical Science, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan.
Exp Neurol. 2003 Dec;184(2):904-11. doi: 10.1016/j.expneurol.2003.08.022.
The ependyma is one of the feasible targets for gene transfer to the brain. Using two different replication-deficient recombinant adenoviral vectors, AdCMVbetaGal or AdRSVIL10, we examined effects of cortical brain ischemia on transgene expression in the ependyma after administration of the vector into the lateral ventricle of spontaneously hypertensive rats (SHR). Expression of the reporter gene lacZ at the lateral ventricle was detected by histochemistry for semiquantitative scoring or by biochemical assay for quantitative analysis. Ependymal cells in the ventricles expressed the transgene as early as 6 h after gene transfer in both sham treatment and ischemia treatment. In the sham treatment, the expression peaked at 12 h and slowly decreased toward day 4 and day 7. However, transgene expressions in the ischemic brain on day 4 and day 7 were significantly higher than sham treatment. In the biochemical assay, beta-galactosidase activity detected on day 4 at the periventricular area of the ischemic group (37 +/- 9 mU/mg protein) was significantly greater than that of the sham group (12 +/- 4, P < 0.01). In the enzyme-linked immunosorbent assay for gene transfer of interleukin-10 (IL-10), IL-10 in the cerebrospinal fluid (CSF) of the ischemic group (11,633 +/- 4322 pg/ml) was significantly greater than that in the sham group (2460 +/- 1486, P < 0.05) on day 5. These results suggest that transgene expression in the exo-focal remote area of ependyma is augmented by cortical ischemia, and the ependyma may be a promising target of gene transfer of brain ischemia.
室管膜是基因转移至脑的可行靶点之一。我们使用两种不同的复制缺陷型重组腺病毒载体AdCMVbetaGal或AdRSVIL10，将载体注入自发性高血压大鼠（SHR）侧脑室后，检测皮质脑缺血对室管膜中转基因表达的影响。通过组织化学进行半定量评分或通过生化测定进行定量分析来检测侧脑室中报告基因lacZ的表达。在假手术组和缺血组中，基因转移后6小时，脑室中的室管膜细胞即表达转基因。在假手术组中，表达在12小时达到峰值，并在第4天和第7天缓慢下降。然而，缺血脑在第4天和第7天的转基因表达明显高于假手术组。在生化测定中，缺血组在第4天在脑室周围区域检测到的β-半乳糖苷酶活性（37±9 mU/mg蛋白）明显高于假手术组（12±4，P<0.01）。在白细胞介素-10（IL-10）基因转移的酶联免疫吸附测定中，缺血组在第5天脑脊液（CSF）中的IL-10（11,633±4322 pg/ml）明显高于假手术组（2460±1486，P<0.05）。这些结果表明，皮质缺血可增强室管膜外灶性远隔区域的转基因表达，室管膜可能是脑缺血基因转移的一个有前景的靶点。
